KR20160081898A - 1,3,4-oxadiazole and 1,3,4-thiadiazole derivatives as immunomodulators - Google Patents

Abstract

The present invention relates to 1,3,4-oxadiazole and 1,3,4-thiadiazole compounds as therapeutic agents capable of inhibiting the prognostic cell death 1 (PD1) signaling pathway. The invention also relates to derivatives of therapeutic agents. The present invention also encompasses the use of such therapeutic agents to treat disorders via immune enhancement, including the inhibition of PD-1, PD-L1, or PD-L2 induced immunosuppressive signals, and therapeutic therapies therewith.

Description

1,3,4-oxadiazole and 1,3,4-thiadiazole derivatives {1,3,4-OXADIAZOLE AND 1,3,4-THIADIAZOLE DERIVATIVES AS IMMUNOMODULATORS}

This application claims the benefit of Indian patent application No. 4012 / CHE / 2013, filed September 6, 2013, which is incorporated herein by reference.

Technical field

The present invention relates to 1,3,4-oxadiazole and 1,3,4-thiadiazole compounds which are therapeutically useful as immunomodulators. The present invention also relates to a pharmaceutical composition comprising a 1,3,4-oxadiazole and a 1,3,4-thiadiazole compound as therapeutic agents.

BACKGROUND OF THE INVENTION

Programmed cell death-1 (PD-1) is a member of the CD28 phase (superfamily) that transmits a negative signal when interacting with its two ligands, PD-L1 or PD-L2. PD-1 and its ligands broadly express and exert a wide range of immunomodulatory roles in T cell activation and resistance to other CD28 members. PD-1 and its ligands weaken infectious immunity and tumor immunity, and are involved in promoting chronic infection and tumor progression. Biological significance of PD-1 and its ligand suggests the therapeutic efficacy of manipulation of the PD-1 pathway for a variety of human diseases (see Ariel Pedoeem et al., Curr Top Microbiol Immunol. (2011); 350: 37).

T-cell activation and dysfunction depend on direct and regulated receptors. Based on their functional consequences, co-signaling molecules can be sub-divided into sub-regulatory factors and co-inhibitors, which are responsible for the priming, growth, differentiation and differentiation of T- Functional maturation is regulated positively and negatively (Li Shi, et al., Journal of Hematology & Oncology 2013, 6:74).

Therapeutic antibodies blocking the prognostic cell death protein-1 (PD-1) immune checkpoint pathway prevent T-cell down-regulation and promote immune responses to cancer. Several PD-I pathway inhibitors have shown robust activity on a variety of ongoing clinical trials (RD Harvey, Clinical Pharmacology & Therapeutics (2014); 962, 214-223).

Predictive cell death-1 (PD-1) is a sub-receptor that is predominantly expressed by T cells. The binding of PD-1 to its ligand, PD-L1 or PD-L2 is very important for the physiological regulation of the immune system. The primary functional role of the PD-1 signaling pathway is the inhibition of autoreactive T cells, which is provided to protect against autoimmune disease. Thus, the elimination of the PD-1 pathway may result in the destruction of immune tolerance, which may ultimately lead to the development of pathogenic autoimmunity. Conversely, tumor cells may occasionally be elected to the PD-1 pathway and escape from the immune surveillance mechanism. Thus, blockade of the PD-1 pathway has become an attractive target for cancer therapies. Current attempts include six agents, PD-1 and PD-L1 targeted neutralizing antibodies or fusion proteins. More than 40 clinical trials are underway to better define the role of PD-1 blockade in various tumor types (see Hyun-Tak Jin et al., Clinical Immunology (Amsterdam, Netherlands) (2014), 153 ), 145-152).

International applications WO 01/14557, WO 02/079499, WO 2002/086083, WO 03/042402, WO 2004/004771, WO 2004/056875, WO2006121168, WO2008156712, WO2010077634, WO2011066389, WO2014055897, WO2014059173, WO2014100079, PD-1 or PD-L1 inhibitory antibodies or fusion proteins.

In addition, international applications WO20011161699, WO2012 / 168944, WO2013144704 and WO2013132317 report peptides or peptidomimetic compounds capable of controlling and / or inhibiting the prognostic cell death 1 (PD1) signaling pathway.

Still, there is a need for more potent and / or better and / or selective immunoregulatory factors in the PD-1 pathway. The present invention provides 1,3,4-oxadiazole and 1,3,4-thiadiazole compounds that are capable of controlling and / or inhibiting the prognostic cell death 1 (PD1) signaling pathway.

SUMMARY OF THE INVENTION

According to the present invention there is provided 1,3,4-oxadiazole and 1,3,4-thiadiazole compounds or pharmaceutically acceptable salts thereof, which are capable of controlling and / or inhibiting the pro-apoptotic 1 (PD1) ≪ / RTI > is provided.

In one aspect, the present invention provides 1,3,4-oxadiazole and 1,3,4-thiadiazole compounds of formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof:

(I)

In the formula (I)

R ₁ is a side chain of an amino acid selected from Ser, Thr, Phe, Ala or Asn;

X is S or O;

R ₂ is hydrogen or -CO-Aaa;

Aaa is an amino acid residue selected from Ser, Asn or Thr, wherein the C-terminus thereof is a free terminal, amidated or esterified;

R ₃ is a side chain of an amino acid selected from Ser, Ala, Glu, Gln, Asn or Asp;

----- is an optional bond;

R ₄ and R ₅ are independently hydrogen or absent.

In a further aspect of the invention this relates to a pharmaceutical composition comprising a compound of formula I, or a pharmaceutically acceptable salt or stereoisomer thereof, and a process for its preparation.

In still another aspect of the present invention, there is provided a pharmaceutical composition comprising 1,3,4-oxadiazole and 1,3,4-thiadiazole of formula (I) capable of controlling and / or inhibiting the intracellular 1 (PD1) Lt; RTI ID = 0.0 > pharmaceutically < / RTI > acceptable salts or stereoisomers thereof.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to the use of 1,3,4-oxadiazole and 1, < RTI ID = 0.0 > 1-oxadiazole < / RTI > as a therapeutic agent useful for the treatment of disorders via immunostimulation involving inhibition of PD- , 3,4-thiadiazole compounds and therapeutic therapies using the same.

Each implementation is provided as a description of the invention and is not provided as a limitation of the invention. Indeed, it will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope or spirit of the invention. For example, features illustrated or described as part of one embodiment may still be used in other embodiments to obtain further embodiments. Accordingly, it is intended that the present invention cover such modifications and variations as come within the scope of the appended claims and their equivalents. Other objects, features, and aspects of the present invention are described in or apparent from the following detailed description. It should be understood by those of ordinary skill in the art that the present invention is illustrative only of exemplary embodiments and should not be construed as limiting the broader aspects of the present invention.

In one embodiment, the invention relates to a compound of formula I, or a pharmaceutically acceptable salt or stereoisomer thereof.

(I)

In the formula (I)

R ₁ is a side chain of an amino acid selected from Ser, Thr, Phe, Ala or Asn;

X is S or O;

R ₂ is hydrogen or -CO-Aaa;

Aaa is an amino acid residue selected from Ser, Asn or Thr, wherein the C-terminus thereof is a free terminal, amidated or esterified;

R ₃ is a side chain of an amino acid selected from Ser, Ala, Glu, Gln, Asn or Asp;

----- is an arbitrary bond;

R ₄ and R ₅ are independently hydrogen or absent.

In still other embodiments, the invention provides a compound of formula IA, or a pharmaceutically acceptable salt or a stereoisomer thereof:

≪ RTI ID = 0.0 &

In the above general formula (IA)

R ₁ is a side chain of an amino acid selected from Ser, Thr, Phe, Ala or Asn;

X is S or O;

R ₂ is hydrogen or -CO-Aaa;

R ₃ is a side chain of an amino acid selected from Ser, Ala, Glu, Gln, Asn or Asp;

Aaa is an amino acid residue selected from Ser, Asn or Thr, wherein the C-terminus thereof is a free terminal, amidated or esterified.

In still other embodiments, the invention provides a compound of formula IB, or a pharmaceutically acceptable salt or stereoisomer thereof:

(IB)

In the above formula (IB)

R ₁ is a side chain of an amino acid selected from Ser, Thr, Phe, Ala or Asn;

R ₃ is a side chain of an amino acid selected from Ser, Ala, Glu, Gln, Asn or Asp;

Aaa is an amino acid residue selected from Ser, Asn or Thr, wherein the C-terminus thereof is a free terminal, amidated or esterified.

In still further embodiments, the invention provides a compound of formula (Ic) or a pharmaceutically acceptable salt or stereoisomer thereof:

≪ EMI ID =

In the above formula (IC)

R ₁ is a side chain of an amino acid selected from Ser, Thr, Phe, Ala or Asn;

R ₃ is a side chain of an amino acid selected from Ser, Ala, Glu, Gln, Asn or Asp;

Aaa is an amino acid residue selected from Ser, Asn or Thr, wherein the C-terminus thereof is a free terminal, amidated or esterified.

In still further embodiments,

R ₁ is a side chain of Ser or Thr;

R ₂ is -CO-Aaa;

Aaa is the amino acid residue Ser or Thr, wherein the C-terminus is free;

R < ₃ > is a side chain of Asn, Gln, Glu or Asp.

The following embodiments are illustrative of the present invention and are not intended to limit the claims to the specific embodiments illustrated.

According to one embodiment, compounds of formula I and formula IA wherein X is O are provided specifically.

According to one embodiment, compounds of formula I and formula IA wherein X is S are specifically provided.

According to one embodiment, compounds of formula I and formula IA wherein R < ₂ > is hydrogen are provided specifically.

According to one embodiment, compounds of formula I are provided wherein R < ₄ > and R < ₅ > are hydrogen.

According to one embodiment, compounds of formula I wherein R < ₄ > and R < ₅ > are absent are provided specifically.

According to still other embodiments, compounds of formula I wherein R < ₂ > is -CO-Ser are provided specifically.

According to still other embodiments, compounds of formula I wherein R < ₂ > is -CO-Thr are provided specifically.

According to still other embodiments, compounds of formula I, formula IA, formula IB and formula IC wherein R < _{1 >} is the side chain of Ser are specifically provided.

According to still other embodiments, compounds of formula I, formula IA, formula IB and formula IC wherein R < ₁ > is the side chain of Thr are provided specifically.

According to still other embodiments, compounds of formula (I), (IA), (IB) and (IC) wherein R ₁ is a side chain of Phe, Ala or Asn are specifically provided.

According to still other embodiments, compounds of formula I, formula IA, formula IB and formula IC wherein R ₃ is the side chain of Asn are specifically provided.

According to still other embodiments, compounds of formula I, formula IA and formula IB wherein R < ₃ > is the side chain of Ser are specifically provided.

According to still other embodiments, compounds of formula I, formula IA and formula IC wherein R < ₃ > is the side chain of Gln are provided specifically.

According to still other embodiments, compounds of formula I, formula IA and formula IC wherein R < ₃ > is the side chain of Glu are provided specifically.

According to still another embodiment, compounds of formula I, formula IA and formula IC wherein R < ₃ > is a side chain of Ala or Asp are provided specifically.

According to still other embodiments, compounds of formula IB and formula IC wherein Aaa is Ser are specifically provided.

According to still other embodiments, compounds of formula (IC) wherein Aaa is Thr are provided specifically.

According to still other embodiments, compounds of formula I, formula IA and formula IB wherein one, more than one or all of the amino acid (s) are D amino acid (s) are specifically provided.

In one embodiment, the specific compounds of formula I, or a pharmaceutically acceptable salt or stereoisomer thereof, without any limitation are listed in Table 1:

Compound No. constitutional formula One

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

And 19

20

The compounds as disclosed in the present invention are formulated for pharmaceutical administration.

In one embodiment, the invention provides a pharmaceutical composition comprising the disclosed compounds, and a pharmaceutically acceptable carrier or diluent.

In other embodiments, the pharmaceutical composition further comprises one or more of an anti-cancer agent, a chemotherapeutic agent, or an anti-proliferative compound.

In one embodiment, the invention provides a compound disclosed herein for use as a medicament.

In another embodiment, the invention provides a compound as disclosed in the present invention for use as a medicament for the treatment of cancer or an infectious disease.

In another embodiment, the present invention provides a method of treating cancer of the bone, of the head or neck, pancreatic cancer, skin cancer, subcutaneous or intraocular malignant melanoma, cervical cancer, ovarian cancer, rectal cancer, cancer of the anal region, gastric cancer, Hodgkin's Disease, non-Hodgkin's lymphoma, esophageal cancer, small bowel cancer, endocrine cancer, thyroid cancer, endometrial carcinoma, endometrial carcinoma, cervical carcinoma, Chronic or acute leukemia, including chronic lymphocytic leukemia, acute lymphoblastic leukemia, chronic leukemia, acute myelogenous leukemia, acute lymphoblastic leukemia, leukemia, cancer of the parathyroid gland, adrenal cancer, soft tissue sarcoma, urethral cancer, , Neoplasms of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal tumors, brain stem glioma, pituitary adenoma, Kaposi < RTI ID = 0.0 > sarcoma, However, There is provided a compound as disclosed in the present invention for use as a medicament for the treatment of cancer of flattened cell, T-cell lymphoma, environmentally induced cancer including those caused by asbestos, and a combination of the above cancers.

In another embodiment, the present invention provides a compound disclosed in the present invention for use in the treatment of cancer.

In another embodiment, the present invention provides a compound disclosed in the present invention for use in the treatment of infectious diseases.

In one embodiment, the invention provides a compound as disclosed in the present invention for use as a medicament for the treatment of a bacterial infectious disease, a viral infectious disease, or a fungal infectious disease.

In one embodiment, the invention provides a method of treating cancer, wherein the method comprises administering to a subject in need thereof an effective amount of a compound of the invention.

In another embodiment, the invention provides a method of modulating an immune response mediated by a PD-I signaling pathway in a subject, comprising administering to the subject a therapeutically effective amount of a compound of the invention, such that the immune response is modulated in the subject ≪ / RTI >

In still other embodiments, the invention provides a method of inhibiting the growth and / or proliferation of tumor cells in a subject, comprising administering to the subject a therapeutically effective amount of a compound of the invention that is capable of inhibiting the prognostic cell death 1 (PD-1) Or < / RTI >

The tumor cells may be selected from the group consisting of bone cancer, cancer of the head or neck, pancreatic cancer, skin cancer, subcutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, gastric cancer, uterine cancer, carcinoma of the fallopian tube, carcinoma of the endometrium Hodgkin's disease, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of the soft tissue, Cancer of the urethra, cancer of the penis, acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, chronic or acute leukemia including chronic lymphocytic leukemia, childhood solid tumor, lymphoid lymphoma, bladder cancer, kidney or uterine cancer Neoplasms of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal tumors, brain stem glioma, pituitary adenoma, Kaposi sarcoma, pancreatic cancer, pancreatic cancer, T-cell lymphoma, asbestosisPlucked environment, including those induced cancer, and cancer including, such as a combination of the arm, and the like.

In yet still further embodiments, the invention provides a method of treating an infectious disease comprising administering to a subject a therapeutically effective amount of a compound of the invention that is capable of inhibiting the prognostic cell death 1 (PD1) signaling pathway, A method of treating an infectious disease in a subject is provided.

Still other further embodiments of the present invention include administering to a subject a therapeutically effective amount of a compound of the invention that is capable of inhibiting the prognostic cell death 1 (PD1) signaling pathway so that the subject is treated for bacterial, viral, and fungal infections And a method for treating bacterial, viral and fungal infections in the subject.

Infectious diseases include pathogenic infections caused by HIV, influenza, herpes, Giardia, malaria, Leishmania, infectious viruses (A, B and C), herpes viruses (e.g. VZV, HSV- I, HAV-6, HSV-II and CMV, Epstein Barr virus), adenovirus, influenza virus, flavivirus, echovirus, (coxsackie virus), cornovirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus But are not limited to, dengue virus, papilloma virus, molluscum virus, poliovirus, rabbit virus, JC virus and arboviral encephalitis virus, But are not limited to, chlamydia, rickettsial bacteria, mycobacteria, staphylococci, streptococcus, pneumonococci, meningococci and conococci ), Klebsiella, Proteus, Serratia, Pseudomonas, etc .; But are not limited to, E. coli, legionella, diphtheria, salmonella, bacilli, cholera, tetanus, botulism, anthrax, Pathogenic infections caused by bacteria, plague, leptospirosis, and Lyme's disease bacteria, fungal Candida (albicans, krusei, glabrata, tropicallis, tropicalis, etc.), Cryptococcus neoformans, Aspergillus (fumigatus, niger, etc.), Mucorales genus Mucor, absidia, rhizophus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis (Paracoccidioides brasiliensis) ), Coccidioides immitis and Histo < RTI ID = 0.0 > Pathogenic infections caused by Histoplasma capsulatum, and pathogenic infections caused by the parasites Entamoeba histolytica, Balantidium coli, Naegleria fowleri, Acanthamoeba sp. Acanthamoeba sp., Giardia lambia, Cryptosporidium sp., Pneumocystis carinii, Plasmodium vivax, Babesia microti, The pathogenicity induced by Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondi, Nippostrongylus brasiliensis, But are not limited to, infection.

The compounds of the present invention can be used as a single drug or as a pharmaceutical composition in which the compound is mixed with various pharmacologically acceptable substances.

The pharmaceutical composition is generally administered by oral or inhalation routes, but may be administered by a parenteral route of administration. In the practice of the present invention, the composition can be administered, for example, orally, by intravenous infusion, topically, intraperitoneally, intracerebrally, or intraspinally. Examples of parenteral administration include, but are not limited to, intra-articular (intra-articular), intravenous, intramuscular, intradermal, intraperitoneal, and subcutaneous routes, and antioxidants, buffers, Aqueous and non-aqueous isotonic sterile injection solutions which may contain solutes which render them isotonic with the blood of the recipient, and aqueous and non-aqueous sterile injection solutions which may include suspending, solubilizing, thickening, stabilizing, Sterile suspensions. Oral administration, parenteral administration, subcutaneous administration and intravenous administration are preferable administration methods.

The dosage of the compounds of the present invention will vary with age, weight, symptoms, therapeutic efficacy, dosage regimen and / or time of treatment. In general, they may be administered orally or by inhalation route in an amount of from 1 mg to 100 mg per dose, once every three days for adults, once every two days, once from once to twice a day, 0.0 > 1 < / RTI > to 24 hours. Since the capacity is affected by various conditions, an amount less than the capacity may sometimes be very sufficient, or in some cases, more capacity may be required.

The compounds of the present invention are useful for the treatment and / or prophylaxis of (1) supplementing and / or improving the prophylactic and / or therapeutic efficacy of the prophylactic and / or therapeutic drugs of the present invention, (2) , Absorption enhancement, dose reduction, and / or (3) reduction of side effects of the prophylactic and / or therapeutic agents of the present invention.

Concomitant medicaments and other medicaments comprising a compound of the present invention may be administered as a combined preparation in which both components are contained in a single formulation or administered in separate formulations. Administration by separate formulations includes simultaneous administration and administration at some time intervals. For administration using some time interval, the compounds of the present invention may be administered first, followed by another drug, or the other compound may be administered first, followed by administering the compound of the present invention. The method of administration of each drug may be the same or different.

The dose of the other drug may be appropriately selected based on the dose used clinically. The compounding ratio of the compound of the present invention and other drugs can be appropriately selected depending on the age and body weight of the subject to be administered, the administration method, the administration time, the disorder to be treated, symptoms, and combinations thereof. For example, the other drug may be used in an amount of 0.01 to 100 parts by mass based on 1 part by mass of the compound of the present invention. The other drug may be a combination of the appropriate ratios with two or more types of any drug. Other drugs that supplement and / or improve the prophylactic and / or therapeutic efficacy of the compounds of the present invention include those that have been discovered, as well as those that have been discovered, on the basis of these mechanisms.

The diseases for which the accompanying use exerts preventive and / or therapeutic effects are not particularly limited. Concomitant medicaments may be used for any disease as long as it supplements and / or enhances the prophylactic and / or therapeutic efficacy of the compounds of the present invention.

The compound (s) of the present invention may be used simultaneously or in the form of a mixture with an existing chemotherapeutic agent. Examples of chemotherapeutic agents are alkylating agents, nitrosoureas, antimetabolites, anticancer antibiotics, vegetable-origin alkaloids, topoisomerase inhibitors, hormone drugs, hormone antagonists, aromatase inhibitors, P-glycoprotein inhibitors, platinum complex derivatives , Other immunotherapeutic drugs and other anti-cancer drugs. It can also be used in the form of a combination with a cancer treatment adjuvant such as a leukopenia (neutropenia) therapeutic agent, a thrombocytopenic treating agent, an anti-vomiting agent and a cancer pain regulating agent.

In one embodiment, the compound (s) of the present invention may be used in combination with other immunomodulators and / or adjuvants, either simultaneously or in admixture. Examples of immunomodulators include various cytokines, vaccines and adjuvants. Examples of these cytokines, vaccines and adjuvants that stimulate the immune response include GM-CSF, M-CSF, G-CSF, interferon-alpha, -beta or γ, IL-1, IL- 12, poly (I: C), including, C, and G _p, are not limited.

In other embodiments, the enhancer is selected from the group consisting of analogs of cyclophosphamide and cyclophosphamide, antitumor inhibitors such as anti-TGF beta and imatinib (Gleevec), paclitaxel, sunitinib (hydroentangel) or other antiangiogenic agents , Aromatase inhibitors such as letrozole, A2a adenosine receptor (A2AR) antagonists, angiogenesis inhibitors, anthracycline, oxaliplatin, doxorubicin, TLR4 antagonists, and IL-18 antagonists.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this subclaim belongs. As used herein, the following definitions apply to promote understanding of the present invention.

As used herein, the term " compound (s) " refers to the compounds described in the present invention.

As used herein, the term " comprises "or" comprising "is generally used to mean including permitting the presence of one or more features or elements.

As used herein, the terms " including "and" include, includes and included "are not limited.

As used herein, the term "amino" refers to a -NH ₂ group. Unless otherwise indicated or listed, all amino groups described or claimed herein may be substituted or unsubstituted.

As used herein, the term "amino acid" refers to an amino acid having L or D stereochemistry at the alpha carbon.

"Pharmaceutically acceptable salts ", especially where the salt is intended to confer improved pharmacokinetic properties in the active ingredient compared to the free form of the active ingredient or any other salt form of the active ingredient used above, Lt; RTI ID = 0.0 > (I) < / RTI > The pharmaceutically acceptable salt form of the active ingredient also encompasses the desired pharmacokinetic properties which have not previously been found and which may even have a positive effect on the pharmacokinetics of the active ingredient in relation to its therapeutic effect in the body, As shown in FIG.

"Pharmaceutically acceptable" means a composition that is generally safe, non-toxic, biologically or otherwise undesirable to produce a pharmaceutical composition, and includes those that are acceptable for veterinary and also human pharmaceutical use do.

The term "stereoisomer " refers to any enantiomer, diastereomer, or geometric isomer of a compound of formula I, whether or not they are chiral or when they have one or more double bonds. When the compounds of formula I and the related formulas are chiral, they may exist in racemic or optically active forms. It may be desirable to use enantiomers because the pharmaceutical activity of the racemates or stereoisomers of the compounds according to the invention may be different. In these cases, the end products or even intermediates can be separated into enantiomeric compounds by chemical or physical scaling used as such in the synthesis or even known to those skilled in the art. In the case of racemic amines, diastereoisomers are formed from the mixture by reaction with optically active solubilizers. Examples of suitable disintegrating agents include R and S of N-protected amino acids such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (such as N-benzoyl proline or N-benzenesulfonyl proline) Or optically active acids such as various optically active camphorsulfonic acids. Chromatographic enantiomeric cleavage using optically active resolving agents (e. G., Dinitrobenzoylphenylglycine, cellulose triacetate or other derivatives of carbohydrates or chiral derivatized methacrylate polymers immobilized on silica gel) It is advantageous.

The term "subject" includes mammals (especially humans) and other animals such as domestic animals (e.g. domestic pets including cats and dogs) and non-domestic animals (e.g. wild animals).

(I) treatment or prevention of a particular disease, disorder or condition, (ii) attenuating, alleviating or eliminating one or more symptoms of a particular disease, disorder or condition, or (iii) Refers to a sufficient amount of the compound (s) of the present invention to prevent or delay the onset of one or more symptoms of the particular disease, disorder or symptom described herein. In the case of cancer, the therapeutically effective amount of the drug may decrease the number of cancer cells; Reduce the size of the cancer and / or; Inhibiting cancer cell infiltration into the peripheral organs (i. E., Stopping to some degree) and / or; Control tumor metastasis (i. E., Slow to some extent and stop running) and / or; To some extent inhibit tumor growth and / or alleviate one or more symptoms associated with cancer to a certain extent. In the case of an infectious disease state, a therapeutically effective amount is an amount sufficient to reduce or alleviate symptoms of an infectious disease, a bacterium, a virus, and an infection caused by the fungus.

Naturally occurring amino acids are identified throughout the specification by the conventional three-letter abbreviations shown in Table 2 below:

Amino acid code designation  3-character code  designation  3-character code Asparagine  Asn  Glutamine  Gln Aspartic acid  Asp  Phenylalanine  Phe Alanine  Ala  Serine  Ser Glutamic acid  Glu  Threonine  Thr

The abbreviations used throughout the specification may be summarized below in the context of their specific meaning.

℃ (degrees Celsius); delta (delta); %(percent); Brine (NaCl solution); CH ₂ Cl ₂ / DCM (dichloromethane); br s (wide single line); Cs ₂ CO ₃ (cesium carbonate); d (double line); DMF (dimethylformamide); DMSO (dimethyl sulfoxide); DMSO-d ₆ (deuterated DMSO); EDC.HCl / EDCI (1- (3-dimethylaminopropyl) -3-carbodiimide hydrochloride); Et ₂ NH (diethylamine); Fmoc (fluorenylmethyloxycarbonyl chloride); g or gr (grams); H or H ₂ (hydrogen); H ₂ O (water); HOBt / HOBT (1-hydroxybenzotriazole); HCl (hydrochloric acid); h or hr (hour); Hz (Hertz); HPLC (high-performance liquid chromatography); I ₂ (iodine); K ₂ CO ₃ (potassium carbonate); LCMS (liquid chromatography mass spectrometer); MeOH (methanol); mmol (millimoles); M (mol); μl (microliter); mL (milliliters); mg (milligrams); m (polylines); MHz (megahertz); MS (ES) (mass spectrometer-electrospray); min. (min); Na (sodium); NaHCO ₃ (sodium bicarbonate); _{NH 2 NH 2 · N 2 O} ( hydrazine hydrate); NMM (N-methylmorpholine); Na ₂ SO ₄ (sodium sulfate); N ₂ (nitrogen); NMR (nuclear magnetic resonance spectroscopy); PD-L1 (prospective cell death-ligand 1); PD-L2 (prospective cell death 1 ligand 2); Preparative-HPLC / preparative HPLC (preparative high-performance liquid chromatography); S (single line); ^t Bu ( ^t -butyl); TEA / Et ₃ N (triethylamine); TLC (thin layer chromatography); THF (tetrahydrofuran); TIPS (triisopropylsilane); TFA / CF ₃ COOH (trifluoroacetic acid); t (triplet); t _R = (retention time); TPP (triphenylmethane); Etc

Experiment

Embodiments of the present invention provide the preparation of compounds of formula I according to the procedures of the following examples, using the appropriate materials. It will be understood by those skilled in the art that the following manufacturing process conditions and known variations of the processes may be used to prepare these compounds. In addition, by using the procedures described in detail, one of ordinary skill in the art can prepare additional compounds of the present invention.

Starting materials are generally commercially available from Sigma-Aldrich, USA or Germany; Chem-Impex, USA; G.L. Biochem and Spectrochem of India.

Purification and Characterization of Compounds

Analytical HPLC Assay: Analytical HPLC was performed using a ZIC HILIC 200 Å column (4.6 mm x 250 mm, 5 μm), flow rate: 1.0 mL / min. The elution conditions used were buffer A: elution by equilibration of the column with 5 mmol of ammonium acetate, buffer B: acetonitrile, 90% buffer B and gradient of 90% to 40% buffer B for 30 min.

Preparative HPLC method: Preparative HPLC was performed using a SeQuant ZIC HILIC 200 Å column (10 mm × 250 mm, 5 μm), flow rate: 5.0 mL / min. The elution conditions used were as follows: Buffer A: 5 mM ammonium acetate (adjusted to pH-4 with acetic acid), buffer B: equilibration of the column with acetonitrile, 90% buffer B and 90% to 40% B elution.

LCMS was performed on an AP1 2000 LC / MS / MS triple quad (Applied bio systems) with an Agilent 1100 series HPLC with G1315 B DAD using a mercury MS column or with a G1315 B DAD Agilent LC / MSD VL with Agilent 1100 Series HPLC A single quad, mercury MS column was used or a Shimadzu LCMS 2020 single quad was used as a Prominence UFLC system with SPD-20 A DAD.

Example 1: Synthesis of Compound (1)

Step 1a:

Potassium carbonate (7.9 g, 57.39 mmol) and methyl iodide (1.3 mL, 21.04 mmol) were added to a solution of compound 1a (5.0 g, 19.13 mmol) in DMF (35 mL) and stirred at room temperature for 2 h. Completeness of the reaction was confirmed by TLC analysis. The reaction mixture was partitioned between water and ethyl acetate. The organic layer was washed with water, brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure to give 5.0 g of compound ( 1b ) (yield: 96.1%). LCMS: 176.1 (M-Boc) ^< ^{+ &} gt ^; .

Step 1b:

Hydrazine hydrate (7.2 mL) was added to a solution of compound ( Ib ) (5.0 g, 18.16 mmol) in methanol (30 mL) and stirred at room temperature for 2 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was evaporated under reduced pressure and the residue obtained was partitioned between water and ethyl acetate. The organic layer was washed with water, brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure to give the compound (1c) of 4.0g (yield: 80.0%). LCMS: 276.3 (M + H) < ⁺ & gt ^; .

Step 1c:

NMM (0.67 ml, 6.52 mmol) was added to a solution of compound 1c (1.2 g, 4.35 mmol), compound 1d (1.43 g, 4.35 mmol), HOBt (0.7 g, 5.22 mmol) and EDC.HCl (0.99 g, 5.22 mmol) in dichloromethane at 0 < 0 > C. The reaction mixture was stirred at room temperature for 12 h. The completeness of the reaction was confirmed by TLC analysis. The reaction was quenched with ice and the precipitated solid was filtered and dried under vacuum to give 2.0 g of pure product ( 1e ) (yield: 83.3%). LCMS: 591.5 (M + Na) < ⁺ & gt ^; .

Step 1d:

Triphenylphosphine (1.38 g, 5.27 mmol) and iodine (1.33 g, 5.27 mmol) were added to a stirred solution of compound 1e (1.5 g, 2.63 mmol) in anhydrous THF (15.0 mL) and DMF Lt; / RTI > After iodine was completely dissolved, Et ₃ N (1.52 mL, 10.54 mmol) was added to the reaction mixture at ice-cold temperature. The reaction mixture was allowed to reach room temperature and stirred for 4 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction was quenched with ice water and extracted with ethyl acetate. The organic layer was washed with saturated sodium thiosulfate and brine solution. The separated organic layer was dried over Na ₂ SO ₄ and evaporated under reduced pressure to give a residue which was purified by silica gel chromatography (eluent: 30% ethyl acetate in hexanes) to give 0.8 g of compound ( 1f ) (Yield: 55%). LCMS: 551.3 (M + H) < ⁺ & gt ^; .

Step 1e:

The Fmoc group was deprotected by adding diethylamine (20.0 mL) to a solution of compound ( 1f) (0.8 g, 1.45 mmol) in CH ₂ Cl ₂ (20.0 mL) at 0 ° C. The reaction was stirred at room temperature for 2 hours. The resulting solution was concentrated in vacuo to give a thick gummy residue. The crude compound was purified by neutral alumina column chromatography (eluent: 2% methanol in chloroform) to give 0.38 g ( 1 g ) of compound (yield: 80.0%). LCMS: 329.4 (M + H) ⁺ .

Step 1f:

The compound was dissolved in DMF (10mL) (1g) ( 0.38g, 1.16mmol), TEA (0.33mL, 2.32mmol) of the compound (1h) for urea bond formation in 0 ℃ To a solution of (0.55g, 1.39mmol) The mixture was added dropwise and the mixture was stirred at room temperature for 2 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction was quenched with ice water and the precipitated solid was filtered off and dried in vacuo to give the crude compound which was further purified by silica gel column chromatography (eluent: 0-35% ethyl acetate in hexanes) to give 0.4 g of The product ( 1i ) was obtained (yield: 59.7%). LCMS: 586.4 (M + H) < ⁺ & gt ^; .

Step 1g:

To a solution of compound ( 1i) (0.4 g, 0.68 mmol) in CH ₂ Cl ₂ (5 mL) was added trifluoroacetic acid (5 mL) and a catalytic amount of triisopropylsilane and stirred at room temperature for 3 hours to give the acid sensitive protective group . The resulting solution was concentrated under nitrogen and the solid material was purified by preparative HPLC method as described under experimental conditions (yield: 0.05 g). LCMS: 318.0 (M + H) < ⁺ >; HPLC: t _R = 10.96 min.

Compound (1h) (NO ₂ -C ₆ H ₄ -OCO-Thr (tBu) -O ^t Bu) Synthesis of:

DCM (25.0mL) solution of 4-nitrophenyl chloroformate H-Thr (tBu) -OtBu ( 5.0g, 21.61mmol) in CH ₂ Cl ₂ (25mL) to a solution of (4.79g, 23.77mmol) TEA (6.2mL , 43.22 mmol) was slowly added at 0 < 0 > C and allowed to stir for 30 minutes. Completion of the reaction was confirmed by TLC analysis. After completion of the reaction, it was diluted with DCM and washed with 1.0 M citric acid followed by 1.0 M sodium carbonate solution. The organic layer was dried over Na ₂ SO ₄ and evaporated under reduced pressure to give crude compound ( 1h ) which was further purified by silica gel column chromatography (eluent: 0-5% ethyl acetate in hexanes) to give 3.0 g of The product ( 1h ) was obtained. ¹ H NMR (CDCl ₃ , 400 MHz) :? 1.17 (s, 9H), 1.28 (d, 3H), 1.50 (s, 9H), 4.11 , ≪ / RTI > 1H), 7.37 (d, 2H), 8.26 (d, 2H).

Example 2: Synthesis of compound (2)

Step 2a:

NMM (1.8 mL, 18.15 mmol) was added to a solution of compound 1c (2.0 g, 7.26 mmol), compound 2d (4.3 g, 7.26 mmol), HOBt (1.17 g, 8.7 mmol) and EDC.HCl (1.66 g, 8.7 mmol) in tetrahydrofuran at 0 < 0 > C. The reaction mixture was stirred at room temperature for 12 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction was quenched with ice and the precipitated solid was filtered and dried under vacuum to give 3.7 g of pure product ( 2e ) (yield: 59.6%). LCMS: 854.4 (M + H) < ⁺ & gt ^; .

Step 2b:

Triphenylphosphine (2.28 g, 8.66 mmol) and iodine (2.2 g, 8.66 mmol) were added to a stirred solution of compound 2e (3.7 g, 4.33 mmol) in anhydrous THF (25.0 mL) and DMF Was added at 0 < 0 > C. After completely dissolving the iodine, Et ₃ N (2.5 mL, 17.32 mmol) was added at the same temperature. The reaction mixture was stirred at room temperature for 4 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction was quenched with ice water and extracted with ethyl acetate. The organic layer was washed with saturated sodium thiosulfate and brine solution. The separated organic layer was dried over Na ₂ SO ₄ and evaporated under reduced pressure, which was further purified by silica gel column chromatography (eluent: 30% ethyl acetate in hexanes) to give 2.0 g of 2f (yield: 55 %). LCMS: 858.4 (M + Na) < ⁺ & gt ^; .

Step 2c:

Diethylamine (30.0 mL) was added to a solution of compound ( 2f) (2.0 g, 1.17 mmol) in CH ₂ Cl ₂ (30.0 mL) at 0 ° C. The reaction mixture was stirred at room temperature for 1 hour. The resulting solution was concentrated in vacuo to give a thick sticky residue. The crude compound was purified by neutral alumina column chromatography (eluent: 2% methanol in chloroform) to give 1.0 g of compound ( 2g ) (yield: 71.4%). LCMS: 614.5 (M + H) < ⁺ & gt ^; .

Step 2d:

Compound 2g (1.0 g, 1.63 mmol) and TEA (0.47 mL, 3.2 mmol) dissolved in DMF (10 mL) was added dropwise at 0 C to a solution of compound ( 1h ) (0.7 g, 1.79 mmol). Thereafter, the reaction mixture was allowed to reach room temperature and stirring was continued for 2 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction was quenched with ice water and the precipitated solids were filtered and dried under vacuum. The obtained crude compound was further purified by silica gel column chromatography (eluent: 0 to 30% ethyl acetate in hexane) to obtain 0.8 g of a product ( 2i ) (yield: 57.1%). LCMS: 871.6 (M + H) < ⁺ & gt ^; .

Step 2e:

To a solution of compound ( 2i) (0.8 g, 0.92 mmol) in CH ₂ Cl ₂ (6 mL) was added trifluoroacetic acid (6 mL) and a catalytic amount of triisopropylsilane and stirred at room temperature for 3 hours. The resulting solution was concentrated under nitrogen and the solid material was purified by preparative HPLC method described under experimental conditions (yield: 0.065 g). HPLC: t _R = 12.01 min; LCMS: 361.34 (M + H) < ⁺ & gt ^; .

Example 3: Synthesis of compound (3)

Step 3a:

Lawesson's reagent (2.85 g, 7.03 mmol) was added to a solution of compound ( 2e ) (4 g, 4.68 mmol) in THF (40 mL) and stirred at 75 ° C for 4 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was evaporated under reduced pressure and the residue obtained was partitioned between ice water and ethyl acetate. It was then washed with a brine solution and the organic layer in NaHCO ₃ solution. Dry the organic layer over Na ₂ SO _4, filtered and evaporated under reduced pressure to give the residue, and this, column chromatography on silica gel: A mixture of 2.7g further purified by (eluent hexanes 0 to 5% ethyl acetate in) ( 3a ) (yield: 67.66%). LCMS: 852.3 (M + H) < ⁺ >

Step 3b:

The Fmoc group on compound ( 3a ) was deprotected by adding diethylamine (3.8 mL) to a solution of compound ( 3a) (1 g, 1.17 mmol) in CH ₂ Cl ₂ (3.8 mL). The reaction mixture was stirred at room temperature for 30 minutes. The resulting solution was concentrated in vacuo to give a thick viscous residue. The crude compound was purified by neutral alumina column chromatography (eluent: 0-50% ethyl acetate in hexanes followed by 0-5% methanol in chloroform) to give 0.62 g of compound ( 3b ). LCMS: 630.5 (M + H) < ⁺ & gt ^; .

Step 3c:

To a solution of compound ( 3b) (0.6 g) in CH ₂ Cl ₂ (7.5 mL) was added trifluoroacetic acid (2.5 mL) and a catalytic amount of triisopropylsilane and stirred at room temperature for 3 hours. The resulting solution was concentrated in vacuo to give 0.13 g of compound ( 3 ) which was purified by preparative HPLC method under laboratory conditions. LCMS: 232.3 (M + H) < ⁺ & gt ^; .

Example 4: Synthesis of compound (4)

Step 4a:

Urea coupling was performed by coupling compound ( 3b ) (0.5 g, 7.9 mmol) in THF (10 mL) with compound ( 4e) (0.34 g, 7.9 mmol) at room temperature. Coupling was initiated by addition of TEA (0.16 g, 15.8 mmol) in THF (10 mL) and the resulting mixture was stirred at room temperature. After 12 hours, THF was evaporated from the reaction mass and partitioned between water and ethyl acetate. Purification (0-50% ethyl acetate in hexanes eluent), the organic layer was washed with water, brine, Na ₂ SO ₄ dried over and evaporated under reduced pressure to give Compound (4a), and this silica gel column chromatography To give 0.45 g of product ( 4a ) (Yield: 61.64%). LCMS: 921.8 (M + H) < ⁺ & gt ^; .

Step 4b:

To a solution of compound ( 4a) (0.55 g) in methanol (20 mL) was added 10% Pd-C (0.15 g) under an inert atmosphere. The mixture was stirred under H ₂ atmosphere for 1 h. Completion of the reaction was confirmed by TLC analysis. Subsequently, the Pd-C catalyst was removed by filtration through a pad of Celite ^® and washed with 20 mL of methanol. Evaporation under reduced pressure gave the combined organic filtrate as a product ( 4b ) separation (yield: 0.42 g, 85.71%). LCMS: 831.5 (M + H) < ⁺ & gt ^; .

Step 4c:

To a solution of compound ( 4b) (0.2 g, 0.3 mmol) in CH ₂ Cl ₂ (5 mL) was added trifluoroacetic acid (5 mL) and a catalytic amount of triisopropylsilane and stirred at room temperature for 3 hours. The resulting solution was concentrated in vacuo and the solid material was purified by preparative HPLC method described under experimental conditions (yield: 0.065 g). HPLC: t _R = 14.1 min .; LCMS: 377.3 (M + H) < ⁺ & gt ^; .

The compound (4e), (NO ₂ -C ₆ H ₄ -OCO-Thr (O ^t Bu) -Bzl, < / RTI > Synthesis of:

100mL of a compound Fmoc-Thr ^(t Bu) in DMF to a solution of -OH (15g, 37.73mmol), which was added and the mixture was cooled to give a _{Cs 2 CO 3 (14.75g, 45.2mmol} ) to a 0 ℃. Benzyl bromide (7.74 g, 45.2 mmol) was added to the cooled reaction mixture, and the solution was stirred at ice-cooling temperature for 30 minutes and then at room temperature for 12 hours. The reaction mixture was concentrated under reduced pressure and diluted with ethyl acetate. The organic layer was washed with water followed by brine solution and dried over Na ₂ SO _4. The filtered solution was concentrated and purified by silica gel column chromatography (eluent: 0-30% ethyl acetate in hexanes) to give 18 g of compound ( 4c ) as a white solid. LCMS: 433.1 (M ^{& lt} ; ^{+ & gt;} Bu) ⁺ , 397.2 (M-OBzl) ^< ^{+ &} gt ^; .

The Fmoc group on compound ( 4c ) (25 g, 51.3 mmol) was treated with diethylamine (100 mL) with stirring at room temperature for 1 hour with compound ( 4d ) (25 g, 51.3 mmol) in CH ₂ Cl ₂ Respectively. The resulting solution was concentrated in vacuo and the thick residue was purified by neutral alumina column chromatography (eluent: 0-50% ethyl acetate in hexanes followed by 0-5% methanol in chloroform) to give 10.6 g of compound ( 4d ) Respectively. LCMS: 266.5 (M + H) < ⁺ & gt ^; .

To a solution of compound ( 4d ) (1.5 g, 5.65 mmol) in CH ₂ Cl ₂ (25 mL) was added TEA (1.14 g, 11.3 mmol) and the solution was stirred at room temperature for 5-10 minutes. To this mixture was added a solution of 4-nitrophenyl chloroformate (1.4 g, 6.78 mmol) in CH ₂ Cl ₂ (10 mL) and the resulting mixture was stirred at room temperature for 12 hours. Completion of the reaction was confirmed by TLC analysis. After completion of the reaction, it was diluted with DCM and washed with 1.0 M sodium sulfate solution followed by 1.0 M sodium carbonate solution. The organic layer was dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to give crude compound ( 4e ) which was further purified by silica gel column chromatography (eluent: 0-20% ethyl acetate in hexanes) 0.7 g of product ( 4e ) was obtained. ^{1 H NMR (DMSO- d 6,} 300 MHz): d 1.04 (s, 9H), 1.16 (d, 3H), 4.11 (m, 1H), 5.11 (m, 3H), 6.91 (d, 2H), 7.40 (m, 5 H), 8.10 (d, 2H), 8.26 (br s, 1 H).

Compounds of the following Table 3 were prepared based on the experimental procedure described above.

The compounds shown in Table 4 below, which can be prepared according to procedures analogous to those described above, using suitable modifications known to one of ordinary skill in the art, are also included in the scope of the present application.

Rescue of mouse spleen cell proliferation in the presence of recombinant PD-L1 / PD-L2:

Recombinant mouse PD-L1 (rm-PDL-1, product no. 1019-B7-100 and R & D Systems) was used as the source of PD-L1.

Requirements:

Mouse spleen cells were harvested from 6-8 week old C57 BL6 mice; RPMI 1640 (GIBCO, product number 11875); DMEM containing high glucose (GIBCO, product number D6429); Fetal bovine serum [Hyclone, product number SH30071.03]; Penicillin (10,000 units / ml) -streptomycin (10,000 μg / ml) liquid (GIBCO, product number 15140-122); MEM Sodium pyruvate solution 100 mM (100x), liquid (GIBCO, Cat. # 11360); Non-essential amino acid (GIBCO, product number 11140); L-glutamine (GIBCO, product number 25030); Anti-CD3 antibody (eBiosciences - 16-0032); Anti-CD28 antibody (eBiosciences - 16-0281); ACK digestion buffer (1 mL) (GIBCO, product number-A10492); Histopark (density -1.083 gm / mL) (SIGMA 10831); Tripplane blue solution (SIGMA-T8154); 2 mL of a Norm Ject Luer Lock syringe - (Sigma 2014-12); 40 [mu] M nylon cell filters (BD FALCON 35230); A blood line calculator (Bright line-SIGMA Z359629); FACS buffer (PBS / 0.1% BSA): phosphate buffered saline (PBS) pH 7.2 (HiMedia TS1006) (SIGMA A7050) containing 0.1% bovine serum albumin (BSA) and sodium azide (SIGMA 08591); 5 mM stock solution of CFSE: The CFSE stock solution was prepared by diluting lyophilized CFSE with 180 μL of dimethyl sulfoxide (DMSO C ₂ H ₆ SO, SIGMA-D-5879) and aliquoting into tubes for further use. The working concentration was titrated from 10 [mu] M to 1 [mu] M. (eBioscience-650850-85); 0.05% trypsin and 0.02% EDTA (SIGMA 59417C); 96-well ELISA plates (Corning CLS3390); BD FACS caliber (E6016); Recombinant mouse B7-H1 / PDL1 Fc chimera, (rm-PD-L1 cat no: 1019-B7-100).

protocol

Spleen cell preparation and culture:

The spleen cells harvested in a 50-mL falcon tube were further treated with 1 mL of ACK digestion buffer for 5 minutes at room temperature. After washing with 9 mL of RPMI complete medium, the cells were resuspended in 3 mL of 1xPBS in a 15 mL tube. The spleen cell suspension, which had been covered with 3 mL of Histopaque, was carefully applied to the bottom of the tube without disturbing it. After centrifugation at 800 xg for 20 minutes at room temperature, the opaque layer of spleen cells was carefully collected without disturbing / mixing the layers. Splenocytes were washed with cold 1xPBS and then used for cell-line assays after total cell count using the trypan blue exclusion method.

Culturing the spleen cells in RPMI complete medium (RPMI + 10% fetal bovine serum + 1mM sodium pyruvate + 10,000 units / mL of penicillin and 10,000μg / mL of streptomycin) and 37 ℃ with 5% CO ₂ in a CO ₂ constant temperature and processor Respectively.

CFSE proliferation assay:

CFSE is a dye that passively diffuses into cells and binds to intracellular proteins. 1x10 < ⁶ > cells / mL of harvested spleen cells was treated with 5 [mu] M CFSE solution in pre-warmed 1xPBS / 0.1% BSA solution at 37 [deg. Excess CFSE was quenched using 5 volumes of ice-cold culture medium on the cells and incubated on ice for 5 minutes. CFSE labeled spleen cells were provided three more times using ice-cold complete RPMI medium. The CFSE labeled 1x10 ⁵ of spleen cells were added to a well (well) containing MDA-MB231 cells (and the 1x10 ⁵ cells cultured in glucose-free DMEM medium) or recombinant human PDL-1 (100ng / mL) and test compounds . Splenocytes were stimulated with anti-mouse CD3 and anti-mouse CD28 antibodies (1 μg / mL, respectively) and the cultures were further incubated with 5% CO ₂ at 37 ° C for 72 h. Cells were harvested and washed three times with ice-cold FACS buffer and% proliferation analyzed by flow cell analysis using 488 nm excitation and 521 nm radiation filters.

Data editing, processing, and reasoning:

The percentage of splenocyte proliferation was analyzed using a cell quest FACS program and the percent structure of splenocyte proliferation by the compound was deduced as the% proliferation value and compared to the% stimulated splenocyte proliferation (positive control) as 100% Standardized and analyzed.

Stimulated splenocytes: spleen cells + anti-CD3 / CD28 stimulation

Background Proliferation: spleen cells + anti-CD3 / CD28 + PD-L1

Compound proliferation: spleen cells + anti-CD3 / CD28 + PD-L1 + compounds

Compound effects were tested by adding the desired concentration of compound to anti-CD3 / CD28 stimulated splenocytes in the presence of ligand (PDL-1).

Claims (21)

Claims 1. A compound of formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof:
(I)

In the formula (I)
R ₁ is a side chain of an amino acid selected from Ser, Thr, Phe, Ala or Asn;
X is S or O;
R ₂ is hydrogen or -CO-Aaa;
Aaa is an amino acid residue selected from Ser, Asn or Thr, wherein the C-terminus thereof is a free terminal, amidated or esterified;
R ₃ is a side chain of an amino acid selected from Ser, Ala, Glu, Gln, Asn or Asp;
----- is an optional bond;
R ₄ and R ₅ are independently hydrogen or absent. 2. A compound of formula IA according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof:
≪ RTI ID = 0.0 &

In the above general formula (IA)
R ₁ is a side chain of an amino acid selected from Ser, Thr, Phe, Ala or Asn;
X is S or O;
R ₂ is hydrogen or -CO-Aaa;
R ₃ is a side chain of an amino acid selected from Ser, Ala, Glu, Gln, Asn or Asp;
Aaa is an amino acid residue selected from Ser, Asn or Thr, wherein the C-terminus thereof is a free terminal, amidated or esterified. 3. A compound according to claim 1 or 2, wherein the compound is of formula IB: < EMI ID = 27.1 > or a pharmaceutically acceptable salt or stereoisomer thereof:
(IB)

In the above formula (IB)
R ₁ is a side chain of an amino acid selected from Ser, Thr, Phe, Ala or Asn;
R ₃ is a side chain of an amino acid selected from Ser, Ala, Glu, Gln, Asn or Asp;
Aaa is an amino acid residue selected from Ser, Asn or Thr, wherein the C-terminus thereof is a free terminal, amidated or esterified. 3. A compound of formula (Ic) or a pharmaceutically acceptable salt or stereoisomer thereof:
≪ EMI ID =

In the above formula (IC)
R ₁ is a side chain of an amino acid selected from Ser, Thr, Phe, Ala or Asn;
R ₃ is a side chain of an amino acid selected from Ser, Ala, Glu, Gln, Asn or Asp;
Aaa is an amino acid residue selected from Ser, Asn or Thr, wherein the C-terminus thereof is a free terminal, amidated or esterified. 3. The compound according to claim 1 or 2, wherein X is O. 3. The compound according to claim 1 or 2, wherein X is S. The method according to claim 1,
R ₁ is a side chain of Ser or Thr;
R ₂ is -CO-Aaa;
Aaa is the amino acid residue Ser or Thr, wherein the C-terminus is free;
R ₃ is a side chain of Asn, Gln, Glu or Asp. 3. The compound according to claim 1 or 2, wherein R < ₂ > is hydrogen. 2. A compound according to claim 1, selected from the group consisting of: or a pharmaceutically acceptable salt or a stereoisomer thereof:

10. A pharmaceutical composition comprising at least one compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier or excipient. 11. A pharmaceutical composition according to claim 10, comprising at least one additional pharmaceutical agent, wherein said additional pharmaceutical agent is an anti-cancer agent, a chemotherapeutic agent, or an antiproliferative compound. 10. A compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt or stereoisomer thereof, for use as a medicament. 10. A compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt or stereoisomer thereof for use as a medicament for the treatment of cancer or infectious diseases. 14. The method of claim 13, wherein the cancer is selected from the group consisting of cancer of the bone, cancer of the head or neck, pancreatic cancer, skin cancer, subcutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, gastric cancer, Hodgkin's Disease, non-Hodgkin's lymphoma, esophageal cancer, small bowel cancer, endocrine cancer, thyroid cancer, endometrial carcinoma, endometrial carcinoma, cervical carcinoma, Chronic or acute leukemia, including chronic lymphocytic leukemia, acute lymphoblastic leukemia, chronic leukemia, acute myelogenous leukemia, acute lymphoblastic leukemia, leukemia, cancer of the parathyroid gland, adrenal cancer, soft tissue sarcoma, urethral cancer, , Neoplasms of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal tumors, brain stem glioma, pituitary adenoma, Kaposi < RTI ID = 0.0 > sarcoma, Plasma, Plate Carcinoma, T- cell lymphoma, causing environmental, including those induced by asbestos cancer, and a compound selected from the group consisting of combinations of said cancers. 14. The compound according to claim 13, wherein the infectious disease is a bacterial infectious disease, a viral infectious disease or a fungal infectious disease. 9. A method of modulating an immune response mediated by a PD-1 signaling pathway in a subject, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 9. 9. A method of inhibiting the growth and / or metastasis of tumor cells in a subject, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 9. 16. The method of claim 15, wherein the tumor cell is a cell of a cancer selected from the group consisting of breast cancer, colon cancer, lung cancer, melanoma, prostate cancer, and kidney cancer. 16. The method according to claim 15, wherein the tumor cell is selected from the group consisting of cancer of the bone, cancer of the head or neck, pancreatic cancer, skin cancer, subcutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, gastric cancer, Hodgkin's Disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, cancer of the endocrine system, cancer of the thyroid gland, endometrial carcinoma of the uterine cervix, carcinoma of the uterine cervix, Chronic or acute leukemia, including acute lymphoblastic leukemia, chronic lymphocytic leukemia, childhood solid tumors, cancer of the parathyroid gland, adrenal cancer, soft tissue sarcoma, cancer of the urethra, cancer of the penis, acute myelogenous leukemia, Neoplasms of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal tumor, brainstem glioma, pituitary adenoma, Kaposi ' s sarcoma, Uid skin The method, flat cell carcinoma, T- cell lymphoma, environmental causes, including those induced by asbestos cancer, and the cancer cells selected from the list consisting of a combination of the arm. 10. A method of treating an infectious disease in a subject, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 9. 11. A method of treating bacterial, viral and fungal infections in a subject, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 9.